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Toxicity to birds

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Endpoint:
long-term toxicity to birds: reproduction test
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No guideline. The effects on embryonic development are investigated through direct application of the substance on the egg shell. A great number of eggs (65 per dose) were tested, which contributes to make the results statistically valid.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
read-across: supporting information
Reference
Endpoint:
long-term toxicity to birds: reproduction test
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No guideline. The effects on embryonic development are investigated through direct application of the substance on the egg shell. A great number of eggs (65 per dose) were tested, which contributes to make the results statistically valid.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline available
Principles of method if other than guideline:
Oil is applied on the surface of fertile eggs. Embryonic survival time, teratogenic potential and growth are evaluated.
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No surrogate test material
Dose method:
other: application onto egg shell
Analytical monitoring:
no
Vehicle:
no
Details on preparation and analysis of diet:
No diet
Test organisms (species):
Anas platyrhynchos
Details on test organisms:
TEST ORGANISM
- Common name: Mallard duck
- Source: Commercial hatcheries: Whistling WIngs, Hanover, Ill., and Truslow Farms, Chestertown, Md.
- Age at test initiation (mean and range, SD): Eggs (72hr of development)
Limit test:
no
Total exposure duration (if not single dose):
18 d
Post exposure observation period:
No post-exposure period
No. of animals per sex per dose and/or stage:
65 eggs per dose. For sex ratio, see results.
Control animals:
yes, concurrent no treatment
Nominal and measured doses / concentrations:
5µL per egg
Details on test conditions:
SUBSTANCE APPLICATION:
5µL of paraffin mixture were applied by microliter pipet to the shell surface just below the air space of upright eggs (72 hrs of development) and permitted to spread freely


NO. OF EGGS
- For negative control: 65
- For treated: 65


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 37.5°C
- Relative humidity (%): 70
Details on examinations and observations:
MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: daily
- Remarks: all eggs were candled daily to determine mortality, and dead embryos were removed and examined.


EMBRYONIC WEIGHT
- Time schedule for examinations: All remaining eggs were opened on the 18th day of development and the embryos were removed and weighed.
- Remarks:


PATHOLOGY
- One half of the embryos were defeathered and placed in Bouin's solution for fixation and subsequent examination of soft tissues by the sectioning technique of Wilson (1965) and by dissection.
- The other half were cleared and staiend with alizarine red S according to the method described by Karnofsky (1965) for squeletal examination.



OTHER:
When removed from the shell, sex-ratios, crown-rump lengths, and beak lengths were recorded.
Details on reproductive parameters:
The following parameters were examined per egg:
- Embryos viable
- Clinical signs of toxicity, abnormalities and mortality


OTHER:
- Embryonic growth: embryo weight, crown-rump length, beak length
- Sex ratio
Reference substance (positive control):
no
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
mortality
Remarks:
Embryonic mortality
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
reproductive parameters
Remarks:
Embryo weight
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
reproductive parameters
Remarks:
Sex ratio
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
other: Development: crown-rump length
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
other: Development: beak length
Repellency factors (if applicable):
Not applicable
Mortality and sub-lethal effects:
For details on mortality and sub-lethal effects see Table no.1.
Effects on reproduction:
Effect on embryo development : see table1
Results with reference substance (positive control):
No reference substance
Further details on results:
No other details
Reported statistics and error estimates:
Embryonic weights, crown-rump lengths, and beak lengths were compared by one-way analysis of variance and Duncan's multiple range test. Survival data, sex distribution and numbers of embryos with one or more defects were compared by squared-chi analysis.
No statistical difference ws found between treated and control eggs for any invetigated parameter.

Table 1. Effects of paraffin mixture (5 µL/egg) on Mallard duck embryos

 

 

Control

Paraffin

Number treated

65

65

Percentage survival (Days 3-18)

97

94

Sex ratio, M:F

45:55

44:56

Embryonic Weight (g)

           Male

15.72 ± 1.78

16.34 ± 1.49

           Female

15.75 ± 2.03

15.98 ± 1.58

           Combined

15.74 ± 1.90

16.14 ±1.54

Crown-rump length (mm)

           Male

84.1 ± 3.7

84.9 ± 4.0

           Female

82.7 ± 3.7

83.4 ± 3.0

           Combined

82.8 ± 3.7

84.1 ± 3.5

Beak length (mm)

           Male

13.2 ± 0.9

13.3 ± 0.5

           Female

13.1 ± 0.6

12.9 ± 0.6

           Combined

13.1 ± 0.7

13.1 ± 0.6

Percentage that were abnormal survivors

4.8

3.3

Validity criteria fulfilled:
not applicable
Conclusions:
The application of paraffin mixture on the shell of Mallard duck egg has no effect on embryo development.
Executive summary:

Paraffin mixture constitued of pentdecane, hexadecane, heptadecane, octadecane, nonadecane, 2,2,4,6,6-pentamethylheptane, 2,2,4,4,6,8,8-heptamethylnonane, 2,6,10,14-tetramethylpentadecane, and decahydronaphthalene, mixed in equal proportions, was applied on Mallard duck eggshell (5µL/egg). Embryonic survival, teratogenic potential and growth were evaluated.

No effect of oil application was noted, neither on embryo survival nor development.

Reason / purpose for cross-reference:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Embryotoxic effects of crude oil in Mallard Ducks and Chicks
Author:
Hoffman, D. J.
Year:
1978
Bibliographic source:
Toxicology and Applied Pharmacology 46: 183-190

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Oil is applied on the surface of fertile eggs. Embryonic survival time, teratogenic potential and growth are evaluated.
GLP compliance:
not specified

Test material

Constituent 1
Reference substance name:
Paraffin mixture
IUPAC Name:
Paraffin mixture
Details on test material:
- Name of test material (as cited in study report): mixture of nine paraffin compounds, Paraffin mixture, Paraffin
- Substance type: Petroleum product
- Physical state: Liquid
- Composition of test material, percentage of components: in equal proportions: pentadecane, hexadecane, heptadecane, octadecane, nonadecane, 2,2,4,6,6-pentamethylheptane, 2,2,4,4,6,8,8,-heptamethylnonane, 2,6,10,14-tetramethylpentadecane, and decahydronaphathalene
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No surrogate test material
Dose method:
other: application onto egg shell
Analytical monitoring:
no
Vehicle:
no
Details on preparation and analysis of diet:
No diet

Test organisms

Test organisms (species):
Anas platyrhynchos
Details on test organisms:
TEST ORGANISM
- Common name: Mallard duck
- Source: Commercial hatcheries: Whistling WIngs, Hanover, Ill., and Truslow Farms, Chestertown, Md.
- Age at test initiation (mean and range, SD): Eggs (72hr of development)

Study design

Limit test:
no
Total exposure duration (if not single dose):
18 d
Post exposure observation period:
No post-exposure period
No. of animals per sex per dose and/or stage:
65 eggs per dose. For sex ratio, see results.
Control animals:
yes, concurrent no treatment
Nominal and measured doses / concentrations:
5µL per egg
Details on test conditions:
SUBSTANCE APPLICATION:
5µL of paraffin mixture were applied by microliter pipet to the shell surface just below the air space of upright eggs (72 hrs of development) and permitted to spread freely


NO. OF EGGS
- For negative control: 65
- For treated: 65


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 37.5°C
- Relative humidity (%): 70

Examinations

Details on examinations and observations:
MORTALITY / CLINICAL SIGNS
- Time schedule for examinations: daily
- Remarks: all eggs were candled daily to determine mortality, and dead embryos were removed and examined.


EMBRYONIC WEIGHT
- Time schedule for examinations: All remaining eggs were opened on the 18th day of development and the embryos were removed and weighed.
- Remarks:


PATHOLOGY
- One half of the embryos were defeathered and placed in Bouin's solution for fixation and subsequent examination of soft tissues by the sectioning technique of Wilson (1965) and by dissection.
- The other half were cleared and staiend with alizarine red S according to the method described by Karnofsky (1965) for squeletal examination.



OTHER:
When removed from the shell, sex-ratios, crown-rump lengths, and beak lengths were recorded.
Details on reproductive parameters:
The following parameters were examined per egg:
- Embryos viable
- Clinical signs of toxicity, abnormalities and mortality


OTHER:
- Embryonic growth: embryo weight, crown-rump length, beak length
- Sex ratio
Reference substance (positive control):
no

Results and discussion

Effect levelsopen allclose all
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
mortality
Remarks:
Embryonic mortality
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
reproductive parameters
Remarks:
Embryo weight
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
reproductive parameters
Remarks:
Sex ratio
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
other: Development: crown-rump length
Duration (if not single dose):
18 d
Dose descriptor:
other: NOELR
Effect level:
5 other: µL/egg
Conc. / dose based on:
test mat.
Basis for effect:
other: Development: beak length
Repellency factors (if applicable):
Not applicable
Mortality and sub-lethal effects:
For details on mortality and sub-lethal effects see Table no.1.
Effects on reproduction:
Effect on embryo development : see table1
Results with reference substance (positive control):
No reference substance
Further details on results:
No other details
Reported statistics and error estimates:
Embryonic weights, crown-rump lengths, and beak lengths were compared by one-way analysis of variance and Duncan's multiple range test. Survival data, sex distribution and numbers of embryos with one or more defects were compared by squared-chi analysis.
No statistical difference ws found between treated and control eggs for any invetigated parameter.

Any other information on results incl. tables

Table 1. Effects of paraffin mixture (5 µL/egg) on Mallard duck embryos

 

 

Control

Paraffin

Number treated

65

65

Percentage survival (Days 3-18)

97

94

Sex ratio, M:F

45:55

44:56

Embryonic Weight (g)

           Male

15.72 ± 1.78

16.34 ± 1.49

           Female

15.75 ± 2.03

15.98 ± 1.58

           Combined

15.74 ± 1.90

16.14 ±1.54

Crown-rump length (mm)

           Male

84.1 ± 3.7

84.9 ± 4.0

           Female

82.7 ± 3.7

83.4 ± 3.0

           Combined

82.8 ± 3.7

84.1 ± 3.5

Beak length (mm)

           Male

13.2 ± 0.9

13.3 ± 0.5

           Female

13.1 ± 0.6

12.9 ± 0.6

           Combined

13.1 ± 0.7

13.1 ± 0.6

Percentage that were abnormal survivors

4.8

3.3

Applicant's summary and conclusion

Validity criteria fulfilled:
not applicable
Conclusions:
The application of paraffin mixture on the shell of Mallard duck egg has no effect on embryo development.
Executive summary:

Paraffin mixture constitued of pentdecane, hexadecane, heptadecane, octadecane, nonadecane, 2,2,4,6,6-pentamethylheptane, 2,2,4,4,6,8,8-heptamethylnonane, 2,6,10,14-tetramethylpentadecane, and decahydronaphthalene, mixed in equal proportions, was applied on Mallard duck eggshell (5µL/egg). Embryonic survival, teratogenic potential and growth were evaluated.

No effect of oil application was noted, neither on embryo survival nor development.